A unique intra-molecular fidelity-modulating mechanism identified in a viral RNA-dependent RNA polymerase

Nucleic Acids Research
Weichi LiuPeng Gong

Abstract

Typically not assisted by proofreading, the RNA-dependent RNA polymerases (RdRPs) encoded by the RNA viruses may need to independently control its fidelity to fulfill virus viability and fitness. However, the precise mechanism by which the RdRP maintains its optimal fidelity level remains largely elusive. By solving 2.1-2.5 Å resolution crystal structures of the classical swine fever virus (CSFV) NS5B, an RdRP with a unique naturally fused N-terminal domain (NTD), we identified high-resolution intra-molecular interactions between the NTD and the RdRP palm domain. In order to dissect possible regulatory functions of NTD, we designed mutations at residues Y471 and E472 to perturb key interactions at the NTD-RdRP interface. When crystallized, some of these NS5B interface mutants maintained the interface, while the others adopted an 'open' conformation that no longer retained the intra-molecular interactions. Data from multiple in vitro RdRP assays indicated that the perturbation of the NTD-RdRP interactions clearly reduced the fidelity level of the RNA synthesis, while the processivity of the NS5B elongation complex was not affected. Collectively, our work demonstrates an explicit and unique mode of polymerase fidelity modulation ...Continue Reading

References

Feb 1, 1992·Current Opinion in Genetics & Development·E Domingo
May 1, 1989·Journal of Virology·D A SteinhauerJ J Holland
Oct 3, 1998·Acta Crystallographica. Section D, Biological Crystallography·A T BrüngerG L Warren
Jun 15, 1999·Virology·T V Pestova, C U Hellen
May 24, 2001·Proceedings of the National Academy of Sciences of the United States of America·S CrottyR Andino
May 20, 2003·Proceedings of the National Academy of Sciences of the United States of America·Julie K Pfeiffer, Karla Kirkegaard
Apr 9, 2004·Proceedings of the National Academy of Sciences of the United States of America·Kyung H ChoiMichael G Rossmann
Aug 6, 2004·The Journal of Biological Chemistry·Cristina Ferrer-OrtaNuria Verdaguer
Dec 2, 2004·Acta Crystallographica. Section D, Biological Crystallography·Paul Emsley, Kevin Cowtan
May 10, 2005·The Journal of Biological Chemistry·Jamie J ArnoldCraig E Cameron
Jun 17, 2006·Bioinformatics·Douglas L Theobald, Deborah S Wuttke
Feb 9, 2007·The Journal of Biological Chemistry·Hélène MaletBruno Canard
Jan 11, 2008·The Journal of Biological Chemistry·Dmitry F ZamyatkinKenneth K-S Ng
Feb 5, 2008·Nature Medicine·Marco VignuzziRaul Andino
Aug 1, 2007·Journal of Applied Crystallography·Airlie J McCoyRandy J Read
Jun 24, 2009·Biochimica Et Biophysica Acta·Linda J Reha-Krantz
Oct 31, 2009·Nucleic Acids Research·Aartjan J W te VelthuisEric J Snijder
Feb 4, 2010·Acta Crystallographica. Section D, Biological Crystallography·Paul D AdamsPeter H Zwart
May 12, 2010·Nucleic Acids Research·Liisa Holm, Päivi Rosenström
Jun 10, 2010·Nature Biotechnology·Adam S LauringRaul Andino
Dec 15, 2010·Proceedings of the National Academy of Sciences of the United States of America·Peng Gong, Olve B Peersen
Aug 3, 2012·Proceedings of the National Academy of Sciences of the United States of America·Nina F GnädigMarco Vignuzzi
Sep 25, 2012·Biochimica Et Biophysica Acta·Xin LiuRoger D Kornberg

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Methods Mentioned

BETA
X-ray
electrophoresis
gel filtration

Software Mentioned

ExPASy ProtParam
ImageJ
DALI
PHASER
DSSP
HKL2000
Phenix
Coot
THESEUS
CNS

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